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Structured illumination microscopy (SIM) is a type of super resolution microscopy technology. By using a structured illumination, we can calculate the information that would be lost through transmission in wide field microscopy and reconstruct images the resolution of which breaks the diffraction limit. It can be achieved at small temporal resolution cost. However, the spatial resolution improvement is still not ideal. This research aims to optimize the construction of SIM.

Calibration is the basis of quantitative measurement in optical tweezers. Multiple calibration methods were developed in past years. However, there is no suitable technologies for in situ optical tweezers calibration in living animals. Specific calibration techniques is greatly needed because the requirement of maintain living status and complexity of micro environment. In this study, a method of scanning detection laser with a piezoelectrical mirror was utilized and the Brownian motion of trapped objects was analyzed. The viscosity factor of the fluidic environment or the dynamic radius of the trapped object were measured besides the stiffness of the trap. Microfluidics is an important technology in biology researches capable for accurately controlling flowing of small volumes of fluid. The application of microfluidics to optical tweezers is also investigated in this study, by which cells can be trapped more steadily in order to mimic the environment inside blood vessels. The study proved the feasibility of optical tweezers calibration and measurement using scanning detection laser, accumulated experience of optical tweezers calibration and measurement, and built foundation for in vivo calibration and measurement.